title: Activity of the efflux pump inhibitor phenylalanine-arginine β-naphthylamide against the AdeFGH pump of Acinetobacter baumannii.
abstract: AdeFGH is a multidrug resistance efflux pump of Acinetobacter baumannii belonging to the resistance-nodulation-cell division (RND) family. Analysis of expression of this pump established it as the most prevalent RND pump in clinical isolates of A. baumannii from Canadian hospitals. In this study, the activity of the efflux pump inhibitor (EPI) phenylalanine-arginine β-naphthylamide (PAβN) against the RND efflux pump AdeFGH of A. baumannii was analysed. The adeFGH operon was cloned under the control of the tac promoter and was introduced in single copy into a surrogate Pseudomonas aeruginosa strain that is devoid of its six native efflux pumps. Isopropyl-β-D-thiogalactopyranoside (IPTG) was used to induce expression of the pump and the sensitivity of strains expressing the adeFGH operon to PAβN was analysed in the surrogate strain and in clinical isolates of A. baumannii by measuring the minimum inhibitory concentrations of trimethoprim, chloramphenicol and clindamycin (substrates of AdeFGH) in the presence and absence of PAβN. PAβN was found to inhibit the ability of the AdeFGH pump to efflux all three antibiotics tested in the surrogate P. aeruginosa strain as well as in the A. baumannii strains. However, in A. baumannii strains the inhibition was more profound for clindamycin than for trimethoprim and chloramphenicol. In addition, these data also support the importance of a strain devoid of efflux pumps for screening of EPIs, since the inhibition of AdeFGH by PAβN was found to be more effective in the surrogate P. aeruginosa strain than in the clinical isolates of A. baumannii.
authors.affiliation: Antimicrobial Resistance Research Group, Applied Bioscience Program, Faculty of Health Sciences, University of Ontario Institute of Technology, 2000 Simcoe Street N, Oshawa, ON, Canada L1H 7K4.
authors.forename: Jenny
authors.initials: J
authors.lastname: Cortez-Cordova
authors.forename: Ayush
authors.initials: A
authors.lastname: Kumar
mesh_terms: D040981:Acinetobacter baumannii / Q000187:drug effects* / Q000235:genetics; D000900:Anti-Bacterial Agents / Q000494:pharmacology; D002701:Chloramphenicol / Q000494:pharmacology; D002981:Clindamycin / Q000494:pharmacology; D003001:Cloning, Molecular; D004151:Dipeptides / Q000494:pharmacology*; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D006801:Humans; D026901:Membrane Transport Proteins / Q000235:genetics / Q000378:metabolism*; D008826:Microbial Sensitivity Tests; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics; D014295:Trimethoprim / Q000494:pharmacology
chemical_list: D000900:Anti-Bacterial Agents; D004151:Dipeptides; D004791:Enzyme Inhibitors; D026901:Membrane Transport Proteins; C419365:phenylalanine arginine beta-naphthylamide; D002981:Clindamycin; D002701:Chloramphenicol; D014295:Trimethoprim